LOCUS       sequence-217657-        7400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vivo voltage imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  AUTHORS   Adam Y, Kim JJ, Lou S, Zhao Y, Xie ME, Brinks D, Wu H, Mostajo-Radji
            MA, Kheifets S, Parot V, Chettih S, Williams KJ, Gmeiner B, Farhi 
            SL, Madisen L, Buchanan EK, Kinsella I, Zhou D, Paninski L, Harvey 
            CD, Zeng H, Arlotta P, Campbell RE, Cohen AE
  TITLE     Voltage imaging and optogenetics reveal behaviour-dependent changes 
            in hippocampal dynamics.
  JOURNAL   Nature. 2019 May 1. pii: 10.1038/s41586-019-1166-7. doi: 
            10.1038/s41586-019-1166-7.
  PUBMED    31043747
REFERENCE   2  (bases 1 to 7400)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2019 May 1. pii: 10.1038/s41586-019-1166-7. doi: 
            10.1038/s41586-019-1166-7."
COMMENT     Sequence Label: YA1627: pAAV_hSyn-Dio-paQuasAr3-s-P2A-CheRiff-s
FEATURES             Location/Qualifiers
     source          1..7400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(984..1010)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1026..1970)
                     /codon_start=1
                     /product="blue light-activated optogenetic actuator derived
                     from Scherffelia dubia channelrhodopsin (Hochbaum et al., 
                     2014)"
                     /label=CheRiff
                     /translation="GGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQC
                     SSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKVI
                     LEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALLV
                     SDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLVL
                     AMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKIH
                     EHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV"
     CDS             complement(1971..2027)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(2253..2963,2964..2966))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2295..2316)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2644..2663
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2903..2924
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2973..3731)
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILASGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     regulatory      3728..3737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3747..3780
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3831..3864
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3889..4477
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3942..3962)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4360..4371)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4531..4550)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4585..4604
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4663..4803
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4663..4792
                     /label=AAV2 ITR
     rep_origin      4878..5333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4965..4984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5175..5196
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5350..5369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5469..5491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5529..5547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5615..5719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5720..5788,5789..6580)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5938..5957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6751..7339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7240..7259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccttaga atcgagtagc ctctggaaaa tcggtggcgc atgaaatgaa gctgtcgata
      841 ctggacatag atcgcatgtc tataacgcct tcagtccgtg ctggaagggg tgcgacaggg
      901 ctgggcatgg atgacatctc caattcctcg ggtggttttg attgtggggc catttcctta
      961 gtgttaagta taggctgact ctgagcgtaa tctggaacat cgtatgggta gctcttggcg
     1021 gccgccactt tatcttcctc tgtcacgaag gccagcacgt ccaccttgcg tcccagaaac
     1081 tgggagctca ctggtctcct gatatcgccg tacataatga tatgctcgtg aattttgatc
     1141 cgcaggaagt gccccagcag tccccaaata ttcttggaca gcaggtcggc gatggtatgg
     1201 ccaatagtgg acccagccac gctcagggtg tgcatgcctt caggcccaaa gatgaacagg
     1261 ccgggaaaca tcagccaact agaatagtac acggcagtca tggccagcac cagtttctta
     1321 cagccgccgg caggcatgat atagtaagac tccacgtaga tgattccagc gttgtagaat
     1381 gtctgggttc catacaccag gccgatacag taaaacagcc acttcaccca cccagtggcc
     1441 agagcggctg tcactcccat gcaaatagtt cccaggtcgg acaccagcag agccattgtc
     1501 cgcttattgt atgcctcact caggccggtg atgttagaca gatgaatcag gatcacggga
     1561 catgtcagca gccattcggc atatctcagc catggggtaa tgttccctcc gtacaggaac
     1621 agcatagcag gactggtgaa ctcgaaatag atctccagaa tcactttgat cagctccacg
     1681 gagcacacgt acacttcctc ccatcccacg ctggccttat aggcgtgcca gccgtagaaa
     1741 gccagaaaca gggcagacag gaacacggca aaccactgga agcccctagc tgtttcctgc
     1801 tcccacaggg ccccatagta gatgtagatg gagctgcact gttcctcata cccatgcacg
     1861 gggtggtagg gtggattcac ttgatatcca gctggggcat ggtactcact gcctccggca
     1921 gaatcgtttc caggtggggc gctgtgagcg tctggagcag gagctccgcc gggaccgggg
     1981 ttttcttcca cgtctcctgc ttgctttaac agagagaagt tcgtggctcc ggagcccact
     2041 tcattctcat aacaaaagaa tctggtggct tcagggaagt ccgtggcaca gctgatgaag
     2101 ctgtcaatgc tggacatgct ccgcatgtcg atgacgccct ccgtgcgtgc gggcagaggg
     2161 gccacggggc tgggcatgct gctcatctcc agctcctctg gaggcttgct ctgcggggcc
     2221 atctccttgg tgttgaggat gggctgggac tgcttgtaca gctcgtccat gccgagagtg
     2281 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     2341 ttgctcaggg cggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     2401 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
     2461 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg
     2521 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     2581 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     2641 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     2701 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     2761 atcaggccgt agccgaaggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     2821 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     2881 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     2941 gtgaacagct cctcgccctt gctcaccgga ggatcggcac cggcactggg ttctggtgcc
     3001 tcggtgtcgc ccagaatagc ccgggatctc aacaggataa agccaaagcc gaccttggca
     3061 gtcacgtcca acaccataaa cagcagagtt tcgatgccca ggcccaccac gccagcgccc
     3121 tcagtgccta tgatccacag gatagggtaa gcggtccaca gcaccaagac cagagctgtc
     3181 agggtgttaa aggtagatgc cacctcgggg ccccgctcct ttgcagcaga tcgcagggat
     3241 gtagccagaa catagagcac cactatcatg caaattgtag agaacaacca ccaactgtat
     3301 ctggctatgg ccgtgtggct caaggctccg atgaggccag tgacgatcat cagggcgtcc
     3361 acacccacca gggtgccgat ggtcacccga tccaccttag cgagaagggc caggtgcagc
     3421 agcagaagtg gggtggtaaa cagccactgg gcgtacctgg cataatagat atccaacatt
     3481 tcgcccccga cggacacctc agtaagcccg ataccaaaga acatagacag atatgcggcg
     3541 gatgcgattc cggaggccag gatagtcaca gcgtaatatt cccgggcatc cttatcggtg
     3601 actccccatc cgcggaccag aaagtagaag gttccaatca gcatcagcag agtgccgatg
     3661 cccagccaca gggtttcagg tctgccgtca cccagtaggt cgtaaccagc ctgcagagcg
     3721 atacttacca tggtggcgga gctagcataa cttcgtataa agtatcctat acgaagttat
     3781 ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga ataacttcgt
     3841 ataatgtatg ctatacgaag ttatgaattc gatatcaagc ttatcgataa tcaacctctg
     3901 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3961 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4021 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4081 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4141 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4201 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4261 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     4321 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4381 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4441 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acccgggcgg
     4501 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4561 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4621 ataagctgca ataaactaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4681 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4741 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4801 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4861 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4921 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     4981 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5041 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5101 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5161 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     5221 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     5281 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     5341 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5401 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5461 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5521 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5581 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5641 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5701 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5761 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5821 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5881 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5941 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6001 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6061 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6121 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6181 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6241 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6301 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6361 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6421 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6481 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6541 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6601 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6661 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6721 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6781 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6841 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6901 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6961 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7021 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7081 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7141 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7201 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7261 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7321 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7381 ctggcctttt gctcacatgt
//
